Structural change and interaction analysis of antimicrobial peptides in membrane conditions

• Project/Area Number: 26440072
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biophysics
• Research Institution: Hokkaido University
• Principal Investigator: AIZAWA TOMOYASU 北海道大学, 先端生命科学研究院, 准教授 (40333596)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
• Keywords: ペプチド / 抗菌ペプチド / 遺伝子組換え / NMR / LPS / 大腸菌 / 酵母 / 微生物 / 抗菌

Outline of Final Research Achievements

Antimicrobial peptides are considered an essential part of the innate immune response of plants, invertebrates and vertebrates as they provide host defenses and can target a wide range of pathogenic microorganisms, including bacteria, fungi, yeast, parasites and viruses. Antimicrobial peptides are thought to kill bacteria by breaking their cell membranes, although the exact mechanisms are still unclear. In this study, we investigated efficient way to prepare isotopically labelled antimicrobial peptides for NMR studies. By using an isotopically labelled antimicrobial peptide, we obtained structural information for LPS-bound form.

Research Products

• [Int'l Joint Research] カルガリー大学(カナダ)
• [Int'l Joint Research] パスツール研究所(フランス)
• [Int'l Joint Research] カルガリー大学(カナダ)
• [Journal Article] Overexpression of Antimicrobial, Anticancer, and Transmembrane Peptides in Escherichia coli through a Calmodulin-Peptide Fusion System. (2017)
  • Author(s): Ishida H, Nguyen LT, Gopal R, Aizawa T, Vogel HJ.
  • Journal Title: J Am Chem Soc Volume: 138 Pages: 11318-26
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Expression, purification and characterization of the recombinant cysteine-rich antimicrobial peptide snakin-1 in Pichia pastoris. (2016)
  • Author(s): Kuddus MR, Rumi F, Tsutsumi M, Takahashi R, Yamano M, Kamiya M, Kikukawa T, Demura M, Aizawa T.
  • Journal Title: Protein Expr Purif. Volume: 122 Pages: 15-22
  • DOI: 10.1016/j.pep.2016.02.002
  • NAID: 120006219912
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Lipopolysaccharide-bound structure of the antimicrobial peptide cecropin P1 determined by nuclear magnetic resonance spectroscopy. (2016)
  • Author(s): Baek MH, Kamiya M, Kushibiki T, Nakazumi T, Tomisawa S, Abe C, Kumaki Y, Kikukawa T, Demura M, Kawano K, Aizawa T.
  • Journal Title: J Pept Sci. Volume: 22 Issue: 4 Pages: 214-221
  • DOI: 10.1002/psc.2865
  • NAID: 120005981524
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Efficient production of a correctly folded mouse α-defensin, cryptdin-4, by refolding during inclusion body solubilization. (2015)
  • Author(s): Tomisawa S, Sato Y, Kamiya M, Kumaki Y, Kikukawa T, Kawano K, Demura M, Nakamura K, Ayabe T, Aizawa T.
  • Journal Title: Protein Expr Purif. Volume: 112 Pages: 21-28
  • DOI: 10.1016/j.pep.2015.04.007
  • NAID: 120005820696
  • Peer Reviewed / Open Access
• [Presentation] Mutation analysis of cryptdin-4, antimicrobial peptide from mouse (2017)
  • Author(s): Rina HIRAMINE, Hiroyuki KUMETA, Yasuhiro KUMAKI, Takashi KIKUKAWA, Makoto DEMURA, Kiminori NAKAMURA, Tokiyoshi AYABE, Tomoyasu AIZAWA
  • Organizer: The 2017 Annual Meeting of the Japan Society for Biosciene, Biotechnology
  • Place of Presentation: 京都女子大学（京都府・京都市）
  • Year and Date: 2017-03-17
• [Presentation] Expression, purification and characterization of the recombinant cysteine-rich antimicrobial peptide snakin-1 derived from potato (2017)
  • Author(s): Megumi Yamano, MD Ruhul Kuddus, Farhana Rumi, Takashi Kikukawa, Hiroyuki Kumeta, Yasuhiro Kumaki, Masakatsu Kamiya, Takashi Kikukawa, Makoto Demura, Tomoyasu Aizawa
  • Organizer: The 2017 Annual Meeting of the Japan Society for Biosciene, Biotechnology
  • Place of Presentation: 京都女子大学（京都府・京都市）
  • Year and Date: 2017-03-17
• [Presentation] ジャガイモ由来抗菌ペプチドsnakin-1の安定的な発現・精製系の構築とNMRによる構造解析 (2016)
  • Author(s): 山野めぐみ、神谷昌克、熊木康裕、菊川峰志、出村誠、相沢智康
  • Organizer: 日本農芸化学会
  • Place of Presentation: 札幌コンベンションセンター（北海道・札幌市）
  • Year and Date: 2016-03-27
• [Presentation] 抗菌ペプチドantibacterial factor-2（ABF-2）が持つ特徴的なC末端側領域の機能的役割 (2016)
  • Author(s): 桑畑佳奈、神谷昌克、菊川峰志、出村誠、相沢智康
  • Organizer: 日本生物物理学会北海道支部例会
  • Place of Presentation: 北海道大学（北海道・札幌市）
  • Year and Date: 2016-03-14
• [Presentation] 組換え抗菌ペプチドの安定生産技術とその解析への利用 (2015)
  • Author(s): 相沢智康
  • Organizer: 第１５回日本蛋白質科学会年会
  • Place of Presentation: あわぎんホール（徳島県・徳島市）
  • Year and Date: 2015-06-24
  • Invited
• [Presentation] Elucidation of influential factor for heterologous productivity of the antimicrobial peptide, cecropin P1 using Escherichia coli (2014)
  • Author(s): Abe, C., Nakazumi, T., Beak, M., Kamiya, M., Kikukawa, T., Kawano, T., Demura, M. Aizawa, T.
  • Organizer: 第51回ペプチド討論会
  • Place of Presentation: 徳島大学（徳島市）
  • Year and Date: 2014-10-22
• [Presentation] 抗菌ペプチドcecropin P1の大腸菌発現系における発現効率に影響を与える要因の解明 (2014)
  • Author(s): 阿部千春, 中住太一, 神谷昌克, 菊川峰志, 河野敬一, 出村誠, 相沢智康
  • Organizer: 日本生物物理学会第52回年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-27
• [Presentation] 抗菌ペプチド human defensin 5のNMRによる多量体形成機構の解析 (2014)
  • Author(s): 橋本革, 冨澤聡, 神谷昌克, 菊川峰志, 熊木康裕, 中村公則, 綾部時芳, 相沢智康, 出村誠
  • Organizer: 日本生物物理学会第52回年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-27
• [Presentation] 再構成無細胞系を用いた抗菌ペプチドの直接発現 (2014)
  • Author(s): 冨澤聡, 神谷昌克, 菊川峰志, 出村誠, 相沢智康
  • Organizer: 日本生物物理学会第52回年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-25
• [Presentation] NMRおよびドッキングによる抗菌ペプチドとリポ多糖の複合体構造解析 (2014)
  • Author(s): 櫛引崇弘, 神谷昌克, 相沢智康, 熊木康裕, 菊川峰志, 水口峰之, 出村誠, 川畑俊一郎, 河野敬一
  • Organizer: 日本生物物理学会第52回年会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-25
• [Presentation] 再構成無細胞系を用いた短鎖ペプチドの直接発現 (2014)
  • Author(s): 冨澤聡, 神谷昌克, 菊川峰志, 出村誠, 相沢智康
  • Organizer: 第14回日本蛋白質科学会年会
  • Place of Presentation: ワークピア横浜・横浜産貿ホール（神奈川県）
  • Year and Date: 2014-06-27